中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (10): 1769-1772.doi: 10.3969/j.issn.1673-8225.2012.10.014

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

人脐带间质干细胞培养上清液可下调肝星状细胞转化生长因子β的表达****★

李明亮，姚志成，钟跃思，林  楠，许瑞云，邓美海   

  1. 中山大学附属第三医院肝胆外科，广东省广州市 510630
  • 收稿日期:2011-10-31 修回日期:2011-12-16 出版日期:2012-03-04 发布日期:2012-03-04
  • 通讯作者: 邓美海，博士，主任医师，中山大学附属第三医院肝胆外科，广东省广州市 510630 drdmh@medmail.com.cn
  • 作者简介:李明亮★，男，1985年生，广东省茂名市人，汉族，中山大学在读硕士，主要从事肝胆外科及间质干细胞治疗肝纤维化的研究。 lmlgz@126.com
  • 基金资助:

    广东省自然基金项目(8151008901000112)，广东省自然基金博士启动项目(10451008901004752)，国家自然青年科学基金项目(81000177，81000674)。

Human umbilical cord stromal stem cells-conditioned medium down-regulated the expression of transforming growth factor beta in rat hepatic stellate cells

Li Ming-liang, Yao Zhi-cheng, Zhong Yue-si, Lin Nan, Xu Rui-yun, Deng Mei-hai   

  1. Department of Hepatobiliary Surgery, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou  510630, Guangdong Province, China
  • Received:2011-10-31 Revised:2011-12-16 Online:2012-03-04 Published:2012-03-04
  • Contact: author: Deng Mei-hai, M.D., Chief physician, Department of Hepatobiliary Surgery, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China drdmh@medmail.com.cn
  • About author:Li Ming-liang★, Studying for master’s degree, Department of Hepatobiliary Surgery, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China lmlgz@126.com
  • Supported by:

     the Natural Science Foundation of Guandong Province, No.8151008901000112*; Doctoral Launched Project, Natural Science Foundation of Guandong Province, No.10451008901004752*; Youth Program of the National Natural Science Foundation of China, No.81000177*, 81000674*

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摘要：

背景：人脐带间质干细胞有望成为治疗肝纤维化新的种子细胞，但目前相关的体外实验报道较少。
目的：探讨人脐带间质干细胞培养液上清对大鼠肝星状细胞纤维化形成生物学活性的影响及可能的作用靶点。
方法：将肝星状细胞以1.0×108 L-1接种于六孔板内，无血清培养24 h后，加入2 mL 脐带间质干细胞培养液上清处理肝星状细胞，设为实验组；对照组为加入等量完全培养基处理肝星状细胞；RT-PCR法检测肝星状细胞内转化生长因子β1、collagen-α1、TIMP-2 mRNA表达；Western blot法检测肝星状细胞内转化生长因子β1蛋白表达。
结果与结论：实验组中肝星状细胞内转化生长因子β1、collagen-α1、TIMP-2 mRNA表达较对照组明显下调，且随处理时间的延长，mRNA表达的下调程度亦随之增强(实验组72 h>实验组48 h>对照组)；实验组肝星状细胞内转化生长因子β1蛋白表达较对照组下调，同时具有时间依赖性(对照组>实验组48 h>实验组72 h，P < 0.05)。提示脐带间质干细胞可抑制肝星状细胞纤维化形成的生物学活性，并可能是通过针对转化生长因子β靶点及其下游基因产物起作用。
关键词：肝星状细胞；脐带间质干细胞；转化生长因子β；培养液上清；体外实验
doi:10.3969/j.issn.1673-8225.2012.10.014

关键词: 肝星状细胞, 脐带间质干细胞, 转化生长因子&beta, 培养液上清, 体外实验

Abstract:

BACKGROUND: Human umbilical cord stromal stem cells are expected to become a new seed cell in the treatment of live cirrhosis. However, few studies of in vitro experiment have been reported.
OBJECTIVE: To explore the effect of human umbilical cord stromal stem cells-conditioned medium (USSC-CM) on the biological activity of fibrosis in rat hepatic stellate cells (HSCs), and the acting target.
METHODS: HSCs were seeded in the six orifice plate in proportion (1.0×105 /mL), then cultured without serum for 24 hours;
2 mL USSC-CM were added to treat HSCs as experimental group; while HSCs were treated with equivalent fresh medium as control group. The expression of transforming growth factor β1 (TGF-β1), collagen-α1 and TIMP-2 mRNA in HSCs were detected by RT-PCR. The expression of TGF-β1 protein was determined by Western blot.
RESULTS AND CONCLUTION: The expression of TGF-β1, collagen-α1 and TIMP-2 mRNA in HSCs were down-regulated in experimental group compared to control group, and the tendency enhanced with time (USSC-CM treated 72 h > USSC-CM treated 48 h > control group). And the expression of TGFβ1 protein in HSCs were down-regulated in experimental group compared to control group and showed time dependent manner (control group > USSC-CM treated 48 h group > USSC-CM treated 72 h group, P < 0.05). Human umbilical cord stromal stem cells can inhibit biological activity of HSCs. And it may carry its point by focusing on the acting target related to TGF-β together with downstream production.

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